title: HIF-prolyl hydroxylase 2 inhibition enhances the efficiency of mesenchymal stem cell-based therapies for the treatment of critical limb ischemia.
abstract: Upregulation of hypoxia-inducible transcription factor-1α (HIF-1α), through prolyl-hydroxylase domain protein (PHD) inhibition, can be thought of as a master switch that coordinates the expression of a wide repertoire of genes involved in regulating vascular growth and remodeling. We aimed to unravel the effect of specific PHD2 isoform silencing in cell-based strategies designed to promote therapeutic revascularization in patients with critical limb ischemia (CLI). PHD2 mRNA levels were upregulated whereas that of HIF-1α were downregulated in blood cells from patients with CLI. We therefore assessed the putative beneficial effects of PHD2 silencing on human bone marrow-derived mesenchymal stem cells (hBM-MSC)-based therapy. PHD2 silencing enhanced hBM-MSC therapeutic effect in an experimental model of CLI in Nude mice, through an upregulation of HIF-1α and its target gene, VEGF-A. In addition, PHD2-transfected hBM-MSC displayed higher protection against apoptosis in vitro and increased rate of survival in the ischemic tissue, as assessed by Fluorescence Molecular Tomography. Cotransfection with HIF-1α or VEGF-A short interfering RNAs fully abrogated the beneficial effect of PHD2 silencing on the proangiogenic capacity of hBM-MSC. We finally investigated the effect of PHD2 inhibition on the revascularization potential of ischemic targeted tissues in the diabetic pathological context. Inhibition of PHD-2 with shRNAs increased postischemic neovascularization in diabetic mice with CLI. This increase was associated with an upregulation of proangiogenic and proarteriogenic factors and was blunted by concomitant silencing of HIF-1α. In conclusion, silencing of PHD2, by the transient upregulation of HIF-1α and its target gene VEGF-A, might improve the efficiency of hBM-MSC-based therapies.
authors.affiliation: INSERM UMRS 970, Fontenay aux Roses, France.
authors.forename: Kiave-Yune
authors.initials: KY
authors.lastname: HoWangYin
authors.forename: Céline
authors.initials: C
authors.lastname: Loinard
authors.forename: Wineke
authors.initials: W
authors.lastname: Bakker
authors.forename: Coralie L
authors.initials: CL
authors.lastname: Guérin
authors.forename: José
authors.initials: J
authors.lastname: Vilar
authors.forename: Clément
authors.initials: C
authors.lastname: d'Audigier
authors.forename: Clément
authors.initials: C
authors.lastname: D'Audigier
authors.forename: Laetitia
authors.initials: L
authors.lastname: Mauge
authors.forename: Patrick
authors.initials: P
authors.lastname: Bruneval
authors.forename: Joseph
authors.initials: J
authors.lastname: Emmerich
authors.forename: Bernard I
authors.initials: BI
authors.lastname: Lévy
authors.forename: Jacques
authors.initials: J
authors.lastname: Pouysségur
authors.forename: David M
authors.initials: DM
authors.lastname: Smadja
authors.forename: Jean-Sébastien
authors.initials: JS
authors.lastname: Silvestre
mesh_terms: D000368:Aged; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D016022:Case-Control Studies; D017690:Cell Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D057510:Endovascular Procedures / Q000379:methods; D006614:Hindlimb / Q000098:blood supply*; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000037:antagonists & inhibitors*; D007511:Ischemia / Q000201:enzymology / Q000628:therapy*; D023821:Limb Salvage / Q000379:methods; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D008875:Middle Aged; D064800:Prolyl-Hydroxylase Inhibitors / Q000627:therapeutic use*; D014162:Transfection
chemical_list: D064800:Prolyl-Hydroxylase Inhibitors; C490190:EGLN1 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases
keywords: Angiogenesis; Growth factor; Hypoxia; Ischemia; Mesenchymal stem cells; Transcription factor